Pyrimidine penicillin



Patented June 15, 1954 PYRIMIDINE PENICILLIN Kunio Yamanaka, Kawamura, and

Hiroshi Yamamoto, Yoshio Shimpei Ito, Tokyo, Japan,

assignors to Nippon Kayaku 00., Limited,

Tokyo, Japan No Drawing. Application June 12, 1953, Serial No. 361,442

Claims priority,

Deceinb 6 Claims.

The present application is a continuation-inpart of application Ser'al No. 256,121, filed November 13, 1951, now abandoned.

The invention relates to crystalline salts 2- amino pyrimidine and penicillin, these salts" having the general formula N=C-R1 wherein Pen. designates an acid penicillin radical, R and R1 designate radicals selected from the group consisting of H, lower alkyl and lower alkoxy, and R2 designates a radical selected from hypodermic and fection.

A further object of the invention is to provide for the production of new and therapeutically useful solid penicillin salts of 2-amino pyrimidines by the precipitation of penicillin from a fermentation broth, an aqueous solution of penicillin or its, easily soluble salts (alkali metal or ammonium salts), or an organic solvent solution of penicillin, in such solvents as ethyl, propyl, butyl or amyl acetate or ether etc., by the addition of an equivalent amount of a 2- amino pyrimidine based upon the penicillin content of the solution.

Other and further objects of thepresent invention will appear from the more detailed description set forth below, it being understood that such detailed description is given by way of illustration and explanation only and not by way of limitation, since various changes therein may be made by those skilled in the art without departing from the scope and spirit of the present invention.

We have found that any member of the 2- amino pyrimidine group (a monoamino pyrimidine group which has the NH2 group in the application Japan er 2, 1950 temperatures.

. Protective action against pneumococcal inpenicillin is added,

ethyl acetate, butyl acetate and amyl acetate and precipitates a crystalline salt which stable and resistant to decomposition in air at normal We have also discovered that the salt of a mineral acid such as hydrochloric acid reacts with these Z-amino pyrimidines andthat a penicillin salt such as of sodium or alkali metal reacts with the Z-amino pyrimidine in. water at cooled or normal crystalline salts similar to these mentioned above. Having investigated these reactions, we have found a method which is suitable for the industrial production of the crystalline penicillin salts as mentioned above.

When these 2-amino pyrimidines are added to the penicillin solution in an organic solvent while it is being stirred at cooled or normal temperatures, it dissolves and then the crystals of the desired salt gradually precipitate. This precipitate is separated and removed. The solvent is removed under reduced pressure from the solvent wet precipitate.

When the 2-amino pyrimidine as above is put into a solution of the same organic solvent and the same result can be brought about.

Further, when a salt of the 2-amino pyrimidine above indicated is added to the aqueous solution of the salt of penicillin such as is mentioned above while being stirred at cooled or normal temperatures, the salt of the 2-amino pyrimidine dissolves and then the crystals of the desired salt are deposited. Said crystals of the salt are separated, and dried under reduced pressure. I

In this case, when the salt of the z amino pyrimidine is added as an aqueous solution, the same result can be obtained.

The products thus obtained are not only effective as medicines but also are pure enough to produce salt of metals such as sodium, potassium, calcium, aluminum etc. of penicillin G.

The 2-amino pyrimidine necessary to carry out this process is a member selected from a group consisting of Z-amino pyrimidine, 2- aminol-methyl pyrimidine, Z-amino--methyl- G-methoxy pyrimidine, 2-amino 4 methyl-6- ethoxy pyrimidine, 2-amino-4,6-dimethoxy pyrimidine, 2-amino-4-methoxy pyrimidine, 2- amino-4,6-dimethyl pyrimidine and 2-amino-4,5- dimethyl-G-methoxy pyrimidine and is not industrially of any diflicult composition. Theretemperatures and precipitate 33 tore, this process is advantageous as a process of producing crystalline penicillin salt.

The potency, melting point, angle of rotation and elemental analysis value of each refined penicillin salt of the Z-amino-pyrimidine group produced as mentioned above are shown in Table I which follows:

Example 3 When respective butyl acetate solutions of the 2-amino pyrimidine group prepared as prescribed TABLE I Oxford Melting R e! 2-amino pyrimidine group Unit Calculated Point, Measured 0.

Z-amino pyrimidine 1, 420 1,383 94 1 2-arnino-4-methyl pyrimidine 1; 3801 1', 340 106: 2' 2-an1ino-4-mcthyl-6-methoxy pyrimidine.-. 1,360 1.254 144 3 2-amino-4-methyl-6-ethoxy pyrimidine 1, 230 1, 220 120 4 2-amino-4-6-dirnethoxy pyrimidine. l, 420 1, 213 130. 5 5: 2-amino-4-methoxy pyrimidine- 1, 130 l, 292 100 6 2amino-4-6-dimethyl pyrimidine l l, 310 1, 298 98 7 2-amino4-5-dimethyl-6-methoxy pyrimidine. l, 260 1, 220 129 8 Analysis Valuesof Elements 4 .Angle of Rotation [a] Calculated Measured reLNo' Methanol Solution 0, Per- H, Per- N, Per- 0, Per: H, Per- N, Percent cent cent cent cent cent Example 1 in the following TablelI are added-while being 40 c. c. of penicillin-butyl acetate solution (contained 3.7 g. of penicillin acid) is cooled with ice under stirring, to which a solution of 1.69 g. of 2-amino-4-methyl pyrimidine dissolved in '70 c. c. of butyl acetate is added. The colorless stirredto the butyl. acetate solutiomot penicillin of the same composition andquantity as. Ex.- ample 1 prepared. andkept at 0? to 15 (1., core responding penicillin salt is obtained; with the results shown in Table II, which follows:

TABLE II v nan y u y'ace corres: p0 e- 2 ammo'pymmdme grow used, g. tate, cc. penicillin fi g gfi composisalt,g non o;

2-amino pyrimidine; 0. 95 40v 4. 0 93" 921 5 2-an1ino-4-methyl-6'meth dine l; 39: 4:5 951 2-amino-4-met dine 1.53 0 4.5 92. 5.- 117 2-amino-4- drmethoxy pyrimidine 1.55 50 4.9 100 130.5 2-amino-4-methoxy'pyrimidine 1.25 '30 4.2 91.5 i 100 2aminor4-6-dimethyl1pyrimidine- 1. 23 40' 4:3 93. 97.5 Z-amino:4-5-dimethyh6-methoxy pyrimldine 1.". 53 .1001 4; 6' 94" 127. 5

Example 2 Asolutionof 1.76 g. of 2-amino-4-methyh6- methoxy-pyrimidine hydrochloride dissolved in 25 c. c. of water is added, with stirring, to a solution of 4 g. of penicillin-sodium salt dissolved in 33 c. c. of water cooled to 0 C.-l0? C.

Vigorous stirring of the resulting mixture at0 C;.5 'CL isfollowed by the formation of colorless crystalline precipitates, which is then separated by filtration, washed with cold water and dried at an atmospheric temperatur e under reduced pressure, whereupon 4.4 g. of 2-amino-4'-methyli- 76 V In the above example, even if? the solvent re In caseamyl acetate is used instead. oibutyl acetate in the above Example 3; inorderfto prepare a; penicillin solution; the same'aquantity as of butyl acetate is usedan'd", inlorder' to prepare solutions of the Z-amino pyrimidine group; a quantity 1.1 times as large as of butyl. acetate is used; in case ethyl acetateis used insteadof butyl acetate, in order to prepare a penicillin solution, the same quantity asof." butyl acetate is used, and, in order. to prepare solutions" of. the 2-amino pyrimidinegroup; a quantity 0.5. times; as large as of butyl'acetatei-isiusem.in case'ether is used instead of butyl-acetate,inordertopre+ pare. a penicillin solution, .the-ssame-quantity as ofbutyl acetate is used, and irr-zordentoprepare solutions. ofv the Zearninmpyrimidinet group; a

7 quantity 1.2 times as large as of butylfaceta'tei isv used, and then the yields are-respectively ap proximate: to those in the 1 case of Example: 3.

'quired to dissolve any member of the 2-amino pyrimidine group is added first to the penicillin solution and then the member of the 2-amino pyrimidine group is added later in a state of crystalline powder, no effects on results are observed.

Example 4 6 eiiects by the change of acid composition are observed.

Below is summarized the utility of 2-amino-4- methyl pyrimidine penicillin and 2-amino-4- methyl-G-methoxy pyrimidine penicillin, produced according to this invention as distinct from other penicillin salts. The above mentioned two An aqueous Solution of hydrochloride of any types of penicillin salts are characterized by the member of the 2 aminO pyrimidine group such fact that they do not have such feeble narcotizaas is shown in the following Table In is sepa tion as of procaine penicillin hitherto extensively rately prepared and is added to an aqueous soluused. but are remarkably httle tome and hygro' tion of penicillin sodium salt of the same comscoplc R H t t t position and quantity as in Example -3 prepared s of 0mm 3/ es s and kept at 0 to C. When the mixture is r (1) Case of oral administration-The penistirred,awhite crystallin precipitate is deposited. cillin salt suspended in 1% C. M. C. (carboxy When the precipitate is separated by filtration, methyl cellulose) solution was administered to washed with cold water and dried at the normal mice by using gastric stylet. The results were obtemperature and reduced pressure, a correspondserved after 24 hours, 48 hours and 5 days.

[+ alive; dead] Mice First Mouse Second Mouse Third Mouse Weight Dosage Weight Dosage Weight Dosage is.) (me) Result (a) (ma) Result (2;) a) Penicillin Salts:

2-arnlno-4-methyl pyrimidine penicillin 12 60 12 so 11 55 2-amino 4 -methyl 6 methoxy pyrimidine penicillin 11 55 11 55 11 as ing penicillin salt of the results as shown in The weight of 5 mg. per g. body weight is equiva- Table III are obtained. Table III follows: lent to 300 g. for 60 kg. of a human body.

TABLE III Percentage Melting uantity uantity Yield of zammo Sihydroo i aqueous corres of weld to pomt pyrimidine group chloride solution penicillin theoretical (decom' g cc salt 8 value, position),

percent 0.

Z-amino pyrimidine l. 32 2. 8 65 92 2-amino-4-methyl pyrimidine l. 45 20 3. 6 82 101 2-amino-4-rnethyl-6-ethoxy pyrimidine 1. 9o 4.3 as 111 2-amino-4-6-dimethoxy pyrimidine... 1. 92 25 4. 5 92 130 2-an1ino-4 methoxy pyrimidine 1. 62 25 4. 1 89 99. 5 2-amino-4-6-diniethyl pyrimidine"... 1. 60 25 4. 1 89 97. 5 2 amino 4 5 dimethyl 6 methoxy pyrimidine 1.90 25 4.5 92 127 In case penicillin potassium salt is used instead (2) Case of hypodermic infection-The peniof the penicillin sodium salt of this example, a cillin salt suspended in sterilized 1% C. M. C. solution with 4.0 g. of penicillin potassium salt solution was injected under the skin on the back dissolved in about c. c. of water is used. In of each mouse. The number of mice which had case penicillin N-ethyl piperidine salt is used tolerated the tests in five days was checked. instead of penicillin sodium salt, a solution with 4.5 g. of penicillin N-ethyl piperidine salt disumber ofAnimals Tested 5 it solved in about 40 c. c. of water is used. Then, results respectively approximate to the above s n syweig t 1mg. 3mg. 5mg. 3mg. were obtained.

Further, in case sulphate is used instead of Penicillin S11E51 2- in -4- iii 1 the hydrochloride of the 2-am1no pyrimidine 2 5 3 1., g ;;3.% ?fifi{ 5 4 1 8 group in the above example, the quantity of Sn 110-42119- i idin 1111 ex... 5 a 1 s phate corresponding to that of hydrochloride in 12 213. rfigfi fifgf 5 2 2 e the above Table III is used as in Table IV, below. i

TABLE Iv Results of hygroscopiczty tests Rates of hygroscopicity in a container satu- Quantityof rated with steam at 20 0.: z-amino pyrimidine group suillpha Rates of Weight Increase W I ht (Percent) aamino imidine 1.44 eg z-aminoieili l pyrimidine 1. 57 Penicillin salts (ma) 2-amino-4-methyl-6-niethoxy pyriini 1.88 24 48 72 2-amino-4-n1ethyl-6-ethoxy pyrimidine 2.02 hours hours hours 2-arnino-4-6-dimethoxy pyrimidine 2.04 7 2-a1nino4-methplg iyrimidiie 2% m04mthy1 pyr m dine 2-amino-4-6- ime yrimi ine n1 e 2-amino-4-5-dimethghg-methoxy pyrimidine 2. 02 penicillin G 303.61 0.033 0.261 0.627

2-an1ino-4-methyl-o-methoxy pyrimidine pencilliu G 299. 87 0.123 0.157 0.213 The results of this case are approximate to P12081118 D 30-228 0-050 0-248 0-963 sameness which irhave .z-t-he :gaabovementioned features show efiective blood levelsiior time; suificient'; for ttherapy tasrfihoW-naxinrthejfollowing when they .;are xza'dministered r-to i human ":bodies.

7 5 Those menicillin :zszirlts .Blood vlevels I (1) .Average values in .u./cc. J of blood levels after 300,000 units of penicillinin oil were-injected into human 'muscles:

Timeinhours 1 3' 12 24 .48

Penicillin Salts:

.r2eamino -4.-methyl rpyrimidine penicil- Klin *2: amino -4-methyl -6 methoxy pyrimi- (line 1; 23 0. 53 "Trace (2) Results of measuring blood levelsrrin the case of administering every three hours (without restriction..:of meals) 200,000 units of 2-amino..4=-

' -methyl-6-rnethcxy pyrimidine penicillin G prep- ..aration .(containing'depos-ited calcium carbonate as ai buffer agent) V ing pneumococcal infection of orallyadministered 2-amino-4-methyl-6-methoxy pyrimidine penicillin which is one of the two types of penicillin salts according to this invention andwhich' has especially low hygroscopicity with .ithose of procaine penicillin and penicillin G-Na salt show that the former is superior to the latteritwo.

113.8 @efiectively used together. J. A. P. -A. 38 544-6. (1949) as regards the produc- -,tion .of para-amino benzoic acid within the body (See literature by the administration of procaine penicillin.) (2) There is no fear of allergy by; procaine. Like results on toxicity, on tolerance,.on-hygroscopicity, on blood levels, on lack of allergy and. on protective efiect against pneumococcal in- :fection' (oral administration) are obtained with the other. 2-amino pyrimidines ,of the tables and. with those as set forth in the general fformula r-r- YenLNHT C :G-Jta 'N='C R wherein Pen. designates. an acid penicillinradical,,R and R1 designate radicals selected .from

the group consistingof H, lower .alkyl andlower alkoxy, and R2 designates a radical selectedffrom the group consisting of H and lower alkyl.

What we claim is:

l; A crystalline salt-of a 2-an1in0 pyrimidine and penicillin wherein the Z-amino pyrimidine is v of the general formula N=OR1 wherein R and' Ri designate radicals selected from'the'group consisting of H, lower alkyl and Jlowersalkoxy, and R2 designates a radical-selected from the: group. consisting: of F H and loweri .alkyl.

2. 2-a-mino-4-methyl pyrimidine penicillin. 3. -2-amino-4-methyl- 0- rnethoxy pyrimidine penicillin.

4.' Z-aminokBdimethyl 6 -i.methoxy pyrimidine penicillin.

5. 2-amino-4 6-dimethoxy pyrimidine penicil- 6.'112-amino .4 methyl '6 -.ethoxy :rpyrimidine penicillin.

PROTECTIVE EFFECT AGAINSTiENEUMOCQCCAL INEE'CTION '(OBAL ADMINISTRATION) I Dos I Observation daysiand Number of Deaths Total Average Penicillin Salts age u of vmg 5 6 '73 9 10 Deaths .;days

'iz-amlnoi-methyhfi-methoxy-py hl igg 1 E, airlinepenicillin. 4 9 100. :2 1 7 5.2 Procainepenicillin r. '50 '1 9 .3.0 25 .2 s .318 Penicillin G-Na 100 2 '1' si 8 --4.0

- ()rganisms for test: Pneumococcus TypeI. .Amountoi inoculated strains: 100 MLD.

Test animals: Groups of 10 mice each weighing to g. were used.

Furthenthe points in respect ofwhich the new penicillin salts are considered to be" superior to procaine penicillin are that:

- (.1) As noantigen against sulfona-mide is produced in" the body after administration, theycan ,Iteferences Cited in thefile of this patent UNITED STATES PATENTS Number Name Date 2,545,962 Lott et a1 '-Mar.20,' 1951 2,585,432 Buckwalter Feb. 12, 1952 

1. A CRYSTALLINE SALT OF A 2-AMINO PYRIMIDINE A PENICILLIN WHEREIN THE 2-AMINO PYRIMIDINE IS OF THE GENERAL FORMULA 